Product containing healthful component and process for preparing the same

ABSTRACT

A product which contains a health-promoting component and a process for preparing such a product, in the present invention, using a pulse crop as a raw material. The product accomplishes health promotion very effectively by means of at least one member of the group consisting of liver function improving constituents, cardiac function improving constituents, anti-inflammatory constituents, antifat constituents, antioxidation consititents, antimutagen constituents, and components which have an intestine-regulating effect in single-stomached animals; and the method produces such a product with good efficiency. The product of the present invention can be used directly “as is”; and alternatively, specified components may be concentrated or may be usefully utilized in applied products which use the product of the present invention as a raw material, e. g., food products, livestock feeds, pet foods or drug raw materials, etc. In miso and soy sauce, since salt is added following the koji preparation step, they are high in salt content and lack all-purpose utility; and in addition, since the aging period following the koji preparation step is extremely long, there are problems in productivity. However, the product of the present invention is easy to manufacture and is also a low-cost product.

TECHNICAL FIELD

The present invention relates to a product containing a healthful orsalutary component and a process for preparing the same. In particular,the product of the present invention is prepared from a pulse crop as astarting material. The term “pulse crop” used herein means leguminouscrops such as soybeans, defatted products and hycopotyls thereof, soyprotein extracts, soy protein isolates and so forth, and the term“product” means products as such (plain products) which are made from apulse crop as a starting material, applied products containing the plainproduct as an ingredient, for example, foods, livestock feeds andaquacultural feeds, cosmetics, diets of pets, and precursory productsfor pharmaceutical preparations.

BACKGROUND ART

In general, soybean, which is one of the pulse crops, containsisoflavone compounds including daidzin, daidzein, genistin andgenistein.

The isoflavone compounds are represented by the following formula andDenotative Table.

Denotative Table R1 R2 daidzin H glucose daidzein H H genistin OHglucose genistein OH H

Of these isoflavone compounds, daidzein is an aglycone of daidzin havingits glucose as a glycosidic saccharide hydrolytically separatedtherefrom, and genistein is an aglycone of genistin having its glucoseas a glycosidic saccharide hydrolytically separated therefrom. Withrespect to the isoflavone compounds, contents thereof and percentagesbetween daidzin and daidzein and between genistin and genistein in adefatted soybean are shown in the following Table 1.

TABLE 1 daidzin daidzein genistin genistein defatted 100 3.2 180 4.2soybean (96.9%) (3.1%) (97.7%) (2.3&) (unit: mg/100 g)

It is understood from Table 1 that, in soybeans, daidzin and genistinare contained in large amounts while daidzein and genistein which areaglycones thereof are contained in smaller amounts.

On the other hand, it has been reported that a glycosidic saccharide ishydrolyzed from an isoflavone compound contained in soybeans to form anaglycone in the course of soy sauce or miso (fermented soybean paste)preparation [see Kihara, K.: Journal of Japan Soy Sauce ResearchInstitute (for SHO-KEN), 16, 5, 190(1990)].

According to this report, although hydrolysis of a glycosidic saccharideproceeds to some extent in cooking of a defatted soybean or in a kojipreparation step (seigiku step), most of the saccharide has alreadyhydrolytically been separated in soy sauce cake or a soybean miso.However, since these are highly salinized, ingesting these in largequantities should be avoided. Accordingly, it is difficult to employ anyprocess for preparing these as a process for preparing a food from apulse crop as a starting material.

Soyasaponins occur in soybeans. For example, in a defatted soybean,soyasaponins occur in an amount of 500 to 700 mg/100 g. It has beenreported that soyasaponins consist of 5 varieties of saponins, i.e.,soya saponin I, soya saponin II, soya saponin III, soya saponin A1 andsoya saponin A2 in the form of a mixture [see Kitagawa et al.: YAKUGAKUZASSHI, 104, 2, 162-168(1984)]. Each of these has glucuronic acid as aglycosidic saccharide, and the aglycone which is such a compound thatthe glycosidic saccharide is removed therefrom includes two varieties ofsoyasapogenol A and soyasapogenol B. In general, however, the aglyconeis present as soyasapogenol B. In a commercially available soybean miso,however, neither soyasapogenol A nor soyasapogenol B was detected.

Further, many reports have been made on pharmacological activities ofvarious constituents contained in pulse crops.

For example, a Pueraria lobata Ohwi (kudzu), which is a type of legume,has been used for a long time as a material for an infusion of Puerariaroot (Puerariae Radix) as an herb medicine. It has been reported thatisoflavone compounds and saponins contained in Pueraria root have liverfunction improving effects (therapeutic activities against hepatopathy)[see “Nohara et al.: Journal of Medical and Pharmaceutical Society (forWAKAN-YAKU),. 5, 408-409(1988”, “Y. Niiho et al.: YAKUGAKU ZASSHI., 109,6, 424-431(1989)” and “Y. Niiho et al.: YAKUGAKU ZASSHI., 110, 8,601-611(1990)”]. More specifically, an alcohol concentration, anacetaldehyde concentration and a concentration of ketones in blood canbe lowered to prevent hangovers, nasty aftereffects of drinking or thelike, and metabolic dysfunction of the liver can be prevented. Further,it has been reported that soyasaponins contained in soybeans have ananti-inflammatory effect, and can reduce serum cholesterol, neutral fatsand phospholipids (cardiac function improving, lipotropic, antidiabetic,antifat, and flesh reducing effects) as well as the above-describedeffect in common with pueraria root [see Ohminami et al.: EIYO TOSHOKURYO., 34, 2, 105-108(1981)]. It is said that although soyasaponinshave the effect of improving functions of a living body as describedabove, soyasaponins in the form of a glycoside can not be absorbed in abody. Accordingly, saponins in the form of a glycoside are notintestinally absorbable until they are decomposed into an aglycone,i.e., soyasapogenol B by enterobacteria having β-glucuronidase effect.It is said that the decomposition takes a lot of time and thus theaglycone has little chance of being absorbed by a small intestine tolead to an extremely low absorption efficiency. On the other hand, theenterobacteria having β-glucuronidase effect is not regarded as usefulenterobacteria. Accordingly, placing no reliance on the decomposition bythe enterobacteria, it is desired that soyasaponins be preliminarilydecomposed in vitro into soyasapogenol B to ingest the latter. The samedesire is true of the soy isoflavone compounds.

Further, soyasapogenol B is a triterpene compound and effective as anendotherine (ET) converter enzyme inhibitor (see Japanese PatentApplication Laid-Open (Kokai) No. H7(1995)-188033) and inhibits anendotherine converter enzyme from exhibiting its function, and hencesoyasapogenol B is effective as a remedy for endotherine (ET)-relateddisorders such as hypertension, cerebrovascular contraction aftersubarachnoid hemorrhage, myocardial infarction, arteriosclerosis, heartfailure, renal failure, asthma, and so forth. Further, the triterpenecompound has a lipase inhibitory effect and a glycerophosphatedehydrogenase inhibitory effect. Accordingly, the triterpene compound iscapable of inhibiting the decomposition of lipids to prevent the lipidsfrom being absorbed in a body, and thus, it is effective for theprevention and treatment of obesity (see Japanese Patent ApplicationLaid-Open (Kokai) Nos. H9(1997)-40689 and H9(1997)-67249) and expectedto improve cardial functions and to reduce flesh. Therefore, it isdesired that soyasaponins be decomposed to obtain soyasapogenol B as anaglycone of saponins.

Moreover, it has been reported that free myo-inositol and the like,which are formed by the decomposition of phytic acid in soybeans, have aliver function improving effect of facilitating lipometabolism in aliver (therapeutic activities against hepatopathy) (see Kubota, H.: FOODCHEMICAL, 8, 83-89 (1987).

A further description will be made on promotiing liver functions. It hasbeen reported that miso soup has an anti-cancer effect, and this isbecause the enzyme activity of a liver is enhanced by the ingestion ofmiso soup [see Kanke, S.: SHOKU NO KAGAKU, 224, 50-54(1996)]. Morespecifically, it is reported that when iron as a carcinogen and oxidantand miso are fed to a rat or mouse, glutathione-s-transferase (GST)activity is increased. GST has been known to combine a carcinogen withglutathione to form a glutathione conjugate, which is excreted from abody. It is said that phase II xenobiotic-metabolizing enzymes toenhance carcinogen detoxication and elimination such as NAD(P)H: quinoneoxidoreductase (DT diaphorase) and aldehyde dehydrogenase, as well asconjugating enzymes such as GST and UDP-glucuronosyl-transferase.

Accordingly, many proposals concerning isoflavone compounds contained insoybeans are disclosed in Japanese Patent Application Laid-Open (Kokai)Nos. S62(1987)-126186, H1(1989)-258669, H5(1993)-170756, and so on.

DISCLOSURE OF THE INVENTION

According to the process described in Japanese Patent ApplicationLaid-Open (Kokai) No. S62(1987)-126186, however, most of the resultingisoflavone compounds are daidzin and genistin which each has aglycosidic saccharide, and aglycones are contained in the resultant insmall amounts. Thus, it is impossible to obtain foods and the like whichhave an excellent pharmacological effect as described above.

The process in Japanese Patent Application Laid-Open (Kokai) No.H1(1989)-258669 comprises hydrolytically separating a glycosidicsaccharide from an isoflavone compound by the action of β-glucosidasewhich is one of the enzymes contained in soybeans. However, aglyconesare formed in a small proportion.

The process described in Japanese Patent Application Laid-Open (Kokai)No. H5(1993)-170756 is one which comprises extractively separatingisoflavone compounds from aglycones of the isoflavone compounds formedin soy sauce cake or soy sauce. Although the aglycones of the isoflavonecompounds are formed in the course of soy sauce preparation in a veryhigh proportion as described above, there is the following disadvantage.That is, the aglycones of the isoflavone compounds are present in soysauce cake because of their insolubility, and soy sauce cake per se doesnot serve as a food. Accordingly, the process cannot be employed as aprocess for preparing a food. Further, aglycones of isoflavone compoundsare also formed in a soybean miso at the initial stage of preparation.However, soybean miso has a problem in that it is a highly salinizedfood and therefore ingesting soybean miso in large quantities should beavoided.

With respect to a soybean miso (mame miso), a rice miso (kome miso),Daitokuji Soy nuggets (Daitokuji-natto: a Japanese fermented soy-food inthe form of nuggets), dried-frozen tofu (Kori-tofu-tofu: a Japanese foodmade of soy milk curds) and yuba (yuba: a Japanese food made of a filmwhich forms on a surface of thick soy milk when the soy milk is gentlyheated) as commercially available foods made from a pulse crop as astarting material, contents of daidzin and daidzein and contents ofgenistin and genistein were comparatively measured. The results are asshown in the following Table 2.

TABLE 2 unit: mg/100 g daidzin daidzein genistin genistein soybean misonot detected 78 not detected 57 rice miso 0.66 21 2.3 20 Daitokuji-nattonot detected 49 not detected 42 dried-frozen tofu 0.95 8.4 8.0 11 yuba110 24 160 17 (detection limit: 0.5 mg/100 g)

It is understood from Table 2 that in the soybean miso, the rice misoand the Daitokuji-natto each of which has been subjected to fermentationtreatment, daidzin and genistin have substantially been hydrolyzed, andin particular, the soybean miso and the Daitokuji-natto contain daidzeinand genistein which are aglycones of daidzin and genistin in largeamounts. However, soybean miso and Daitokuji-natto should not beingested in large amounts because of their high salt contents. It isalso understood that in the dried-frozen tofu, each of daidzin,daidzein, genistin and genistein is contained in a small amount. It isfurther understood that similarly to the defatted soybean describedabove, the yuba contains daidzin and genistin in large amounts anddaidzein and genistein which are aglycones thereof in small amounts.

Furthermore, when we look at soybean food products of other countries,“tempeh,” which is a traditional fermented soybean food product ofIndonesia, is a product prepared by fermentation using filiform bacteriaof the genus Rhizopus following fermentation by lactic acid bacteriawithout the addition of salt. This product is used as a raw material forvarious types of side dishes; however, since absolutely no hydrolysisprocess is performed on the products produced by the koji preparationprocess following said koji preparation treatment as in the presentinvention, it has been reported together with the data shown in FIGS.4(a) through 4(c) that the rate of aglyconization in which theisoflavone compounds daidzin and genistin contained in soybeans areconverted into daidzein and genistein (which are aglycones from whichthe respective glycosides have been separated) is extremely low comparedto that of the above-described miso, etc. (Esaki, H. et al.: ACSSymposium Series 546 “Food Phytochemical for Cancer Prevention I” (1994)353-360). Accordingly, aglycones constituting health-promotingcomponents cannot be produced more efficiently and in higherconcentrations by means of the “tempeh”.

With respect to a soybean miso (mame miso), a rice miso (kome miso),Daitokuji Soy nuggets (Daitokuji-natto: a Japanese fermented soy-food inthe form of nuggets), dried-frozen tofu (Kori-tofu, tofu: a Japanesefood made of soy milk curds) and yuba (yuba: a Japanese food made of afilm which forms on a surface of thick soy milk when the soy milk isgently heated) as commercially available foods made from a pulse crop asa starting material, phytic acid contents were measured. The results areas shown in Table 3 below.

TABLE 3 Units: mg/100 g Phytic Acid Soybean miso Not detected Rice misoNot detected Daitokuji natto Not detected Dried-frozen tofu 506 Yuba 361Detection limit: 5 mg/100 g

It is seen from this Table 3 that phytic acid is almost completelybroken down in soybean miso, rice miso and Daitokuji natto, which aresubjected to fermentation treatments. However, soybean miso, rice misoand Daitokuji natto have high salt concentrations, and cannot be eatenin large amounts. Furthermore, it is seen that the phytic acid contentis large in the case of frozen tofu and dried tofu, so that myo-inositolcannot be digested and absorbed.

Furthermore, in regard to other pharmacological actions, soybeans inparticular (among various types of beans) consist chiefly of protein,and miso and soy sauce are famous as food products in which this proteinis broken down into peptides and amino acids by the protease of kojiorganisms. Moreover, in such miso and soy sauce, it is claimed that achemical reaction (Maillard reaction) takes place with carbohydrates andfats which are components of the same soybeans as a result of a drop inthe molecular weight of the protein, so that browning and fragrantaromas (aroma of ripening) are produced. In recent research concerningsuch Maillard reaction products, it has been reported that intermediateAmadori rearrangement products and brown melanoidin or AGE (advancedglycation end product), which are final products, are produced, and thatthese components are anti-mutagenic substances (Hiromich Kato: Miso noKagaku to Gijutsu, Vol. 41, No. 2, 40-48 (1993)). However, the followingproblem arises: i e., miso and soy sauce are food products with a highsalt content and cannot be eaten in large amounts.

If it were possible to ingest large amounts of a food product containinglarge amounts of products including health-promoting components with asuperior pharmacological effect such as those described above, it wouldbe possible to enjoy a diet exhibiting superior effects in terms ofhealth maintenance in humans. Conventionally, however, there has been nofood product which satisfies such a demand.

In particular, components which latently contain health-promotingcomponents such as the above-described isoflavones, saponins or phyticacid, etc., either contain glycosides as in isoflavones and saponins, orform chelate bonds as in phytic acid. Accordingly, such components tendnot to be absorbed by single-stomached animals including human beings orthe gastrointestinal digestive tracts of animals; and hepatic function,etc., cannot be improved regardless of the amounts of such componentsthat are ingested.

Conventionally, furthermore, it has been impossible to convertisoflavones, saponins or phytic acid, etc. into forms that are easilydigested and absorbed.

The present invention has been made in light of the these points, andthe object is to provide a product containing a health-promotingcomponent that includes at least one member of the group consisting ofliver function improving constituents, cardiac function improvingconstituents, anti-inflammatory constituents, antifat constituents,antioxidation consititents and antimutagen constituents; to provideapplied products that contain the above-described product, as aningredient, such as foods which can be ingested in a large amount,livestock feeds, diets of pets and precursory products forpharmaceutical preparations; and further to provide a process forreadily preparing the above-described product in a large amount at a lowcost.

Another object of the present invention is to provide a process forpreparing a product that contains, in addition to the above-describedhealth-promoting component, a health-promoting component which alsocontains large quantities of bacteria that have an intestine-regulatingeffect (hereafter referred to as “intestine-regulating bacteria”) insingle-stomached animals. Here, the term “single-stomached animals”refers to animals which do not have a ruminant stomach such as humanbeings, pets such as dogs and cats, etc., livestock such as horses andpigs, etc., and birds, etc.

In order to accomplish the object, the product of the present inventioncontaining the health-promoting component claimed in claim 1 ischaracterized in that: the product contains a health-promoting componentproduced by inoculating koji mold on a pulse crop to effect kojipreparation, and then hydrolyzing the products produced by this kojipreparation treatment, and the product contains a health-promotingcomponent consisting of intestine-regulating bacteria added to the beansand grown during the period extending from the inoculation of the kojimold to the completion of the hydrolysis.

Furthermore, the process of the present invention for preparing aproduct that contains the health-promoting component claimed in claim 6is characterized in that: the health-promoting component is produced byinoculating koji mold on a pulse crop to effect koji preparation, andthen hydrolyzing (through the addition of water) the products producedby this koji preparation treatment, and the product that contains thehealth-promoting component is produced by adding intestine-regulatingbacteria which constitute a health-promoting component to the beans andgrowing the bacteria during the period extending from the inoculation ofthe koji mold to the completion of the hydrolysis.

The above-described intestine-regulating bacteria are characterized inthat they are added at the same time as the inoculation of the koji moldand consist of lactic acid bacteria which have an intestine-regulatingeffect in single-stomached animals. These intestine-regulating bacteria,and especially lactic acid bacteria, have a good compatibility with thekoji mold used in koji preparation, and also propagate very well ontheir own without interfering with the propagation of the koji mold, sothat production efficiency is also superior.

More specifically, koji mold have the property of being able to coexistcompatibly with lactic acid bacteria, etc. Accordingly, in the presentinvention, an effect in which koji mold and lactic acid bacteria arecaused to be co-present on a substrate consisting of beans (via aprocess involving the co-presence and co-growth of koji mold and lacticacid bacteria) is utilized to produce a food product having an object ofhealth maintenance of intestine-regulation functions, etc. in humans andanimals.

Yogurt may be cited as an example of a healthy food product utilizinglactic acid bacteria; however, yogurt is not a food product whichutilizes koji mold.

The fact that koji mold can co-exist and be co-grown compatibly withlactic acid bacteria, etc., has also been utilized in the past; however,this utilization has been limited to the production of refined sake,shochu, miso and soy sauce, etc., as fermented products utilizing kojimold.

First, in refined sake, lactic acid bacteria are added in order to causethe growth of large quantities of only sake-refining yeast which has astrong activity in the initial stage. This process is called“yamahai-moto”, a traditional Japanese term; here, lactic acid bacteriaare naturally admixed and these lactic acid bacteria produce lacticacid, so that an action, which suppresses the propagation of organisms(film yeast, wild yeast, etc.) that are a problem in the manufacture ofhigh-quality refined sake, is utilized. Today, a process in which lacticacid is added (“tokujo moto” or rapid fermentation) has also beendeveloped.

Meanwhile, in the manufacture of sake, lactic acid bacteria known as“hiochii” bacteria which grow with a preference for alcohol causechanges such as cloudiness, an increase in acid and changes infragrance, etc., and are therefore rather considered to be a problem. Ithas been found that such “hiochii” bacteria are lactic acid bacteriathat require mevalonic acid. It has been confirmed that such mevalonicacid is caused to accumulate with the propagation of organisms in thekoji preparation step by koji mold such as Aspergillus oryzae, etc.Incidentally, it is claimed that lactic acid bacteria that require thismevalonic acid commonly include bacteria that produce bacteriocin.

Furthermore, in miso and soy sauce, lactic acid bacteria are used as acountermeasure to inhibit the growth of undesirable organisms in theaging process.

Thus, products utilizing koji mold are respective products obtained viaprocesses involving the co-presence and co-growth of koji mold andlactic acid bacteria or yeast. However, there has been no food productconsisting of a fermented food having an object of health maintenance(e. g., intestine-regulating action, etc.) in humans and animalsobtained by utilizing an effect which causes the co-presence andco-growth of koji mold and lactic acid bacteria on substrate grains asin the present invention.

The above-described health-promoting component produced by the kojipreparation treatment and hydrolysis treatment are characterized so asto be comprised of at least one member of the group consisting of liverfunction improving constituents, cardiac function improvingconstituents, anti-inflammatory constituents, antifat constituents,antioxidation consititents and antimutagen constituents; and it isfurther characterized in that said component is formed so as to beabsorbed by the digestive tract.

According to the process of the present invention, a health-promotingcomponent can be produced in a product using beans as a raw material bymeans of a koji preparation treatment and a hydrolysis treatment.Furthermore, intestine-regulating bacteria which are added following theinitiation of the koji preparation treatment but prior to the completionof the hydrolysis treatment can be propagated, so that thehealth-promoting component consisting of such intestine-regulatingbacteria can also be included in large amounts.

In concrete terms, proteins, isoflavones, saponins and phytic acid,etc., are respectively converted into the health-promoting componentsuch as peptides, isoflavone aglycones, saponin aglycones, myo-inositoland Maillard reaction products, etc., by performing a koji preparationtreatment and a hydrolysis treatment on beans used as a raw material,and are thus converted into a state in which these components can bevery easily absorbed by the digestive tracts of single-stomachedanimals, etc. Using the product of the present invention which has sucha health-promoting component based on the above-described kojipreparation treatment and hydrolysis treatment, health promotion bymeans of at least one member of the group consisting of liver functionimproving constituents, cardiac function improving constituents,anti-inflammatory constituents, antifat constituents, antioxidationconsititents and antimutagen constituents can be accomplished veryeffectively.

In the present invention, furthermore, intestine-regulating bacteriawhich are added following the initiation of the koji preparationtreatment but prior to the completion of the hydrolysis treatment can bepropagated very favorably, so that a health-promoting componentconsisting of such intestine-regulating bacteria can be included inlarge amounts in the final product, and can thus be used for intestinalregulation in single-stomached animals.

In particular, if intestine-regulating bacteria are added simultaneouslywith the inoculation of koji mold, these intestine-regulating bacteriawill be propagated in large amounts in the process up to the completionof hydrolysis, so that the propagation of other undesirable organismscan be inhibited.

Furthermore, by the use of lactic acid bacteria which have anintestine-regulation action in single-stomached animals as theabove-described intestine-regulating bacteria, an intestine-regulatingaction can be reliably manifested in single-stomached animals, and thepropagation of other undesirable bacteria can be reliably inhibited bythe lactic acid bacteria, which are acid-producing bacteria.

The product thus prepared can be utilized directly, and it also can beutilized in applied products which use this product as a raw material,e. g., food products, livestock feeds, pet foods or drug raw materials,etc., thus being superior in terms of all-purpose utility. Miso and soysauce are foods with a high salt content since salt is added followingthe koji preparation step, thus having less all-purpose utility;moreover, the aging period of such food products following the kojipreparation step is extremely long, thus having problems inproductivity. To the contrary, according to the present invention, theproduct is easy to manufacture and realizes a saving in costs.

Since the product and preparation process of the present invention areconstructed and act as described above, a health-promoting component canbe produced in a product using beans as a raw material, by means of akoji preparation treatment and a hydrolysis treatment. In concreteterms, peptides, isoflavones, saponins and phytic acid, etc. arerespectively converted into a health-promoting component such asisoflavone aglycones, saponin aglycones, myo-inositol and Maillardreaction products, etc. and are thus converted into a state which can bevery easily absorbed by the digestive tracts of single-stomachedanimals. In the present invention, furthermore, the intestine-regulatingbacteria added following the initiation of the above-described kojipreparation treatment but prior to the completion of the hydrolysistreatment propagate so that a health-promoting component consisting ofsuch intestine-regulating bacteria can be included in the product inlarge amounts, thus making it possible to achieve anintestine-regulating effect in the intestines of single-stomachedanimals. By using the product of the present invention which containssuch a health-promoting component, health promotion by means of at leastone member of the group consisting of liver function improvingconstituents, cardiac function improving constituents, anti-inflammatoryconstituents, antifat constituents, antioxidation consititents andantimutagen constituents and components which have anintestine-regulating effect in single-stomached animals can beaccomplished very effectively. This product can be used directly “asis”, or the efficacy can be strengthened by extracting and concentratingthe isoflavone aglycones, saponin aglycones, myo-inositol or Maillardreaction products, etc; in addition, this product can also be utilizedin applied products which use this product as a raw material, e. g.,food products, livestock feeds, pet foods or drug raw materials, etc.and is thus superior in terms of all-purpose utility. Furthermore, misoand soy sauce are foods with a high salt content, since salt is addedfollowing the koji preparation step, so that such food products lackall-purpose utility; moreover, the aging period of such food productsfollowing the koji preparation step is extremely long, so that there areproblems in productivity. However, the product of the present inventionis easy to manufacture at a low-cost.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a process diagram which illustrates a first embodiment of theprocess of the present invention for preparing a product containing ahealth-promoting component.

FIG. 2 is a graph which illustrates the temperature characteristics ofthe mixture accompanying the progression of koji preparation time in thepresent invention.

FIG. 3 is a characteristic diagram obtained by means of an AOM test incases where a product prepared by the process of the present inventionand other substances were used as samples.

FIG. 4 is a characteristic diagram which shows changes in the isoflavonecontent during fermentation in natto, tempeh and miso.

BEST MODE TO CARRY OUT THE INVENTION

Below, embodiments of the present invention will be described withreference to the attached figures.

FIG. 1 is a process diagram which illustrates one embodiment of theprocess of the present invention for preparing a product containing ahealth-promoting component, in which latent health-promoting componentscontained in soybean meal, a type of soybeans, are produced.

To describe the process according to FIG. 1, the soybean meal is firstcooked. The propagation of koji mold is facilitated by this cooking.This cooking of the soybean meal may be performed either continuously orin a batch process according to the object of preparation, etc.

Then, when this cooking is completed, the soybean meal is temporarilycooled, and the moisture content in the soybean meal is adjusted to anamount (e. g., 40 wt %) that allows the propagation of koji mold.

In cases where defatted soybean meal, etc. is used as a raw material,this cooking process may be omitted.

The process of the present invention is performed as described belowusing soybean meal whose moisture content has thus been adjusted.

More specifically, koji starter consisting of koji mold is added at aprescribed weight ratio to the simple soybean meal following thecompletion of cooking; furthermore, lactic acid bacteria constituting atype of intestine-regulating bacteria are added, and these threeingredients are mixed until a uniform mixture is obtained.

Afterward, the mixture is placed in a koji preparation device and heldfor a prescribed period of time with the initial temperature set atapproximately 28 to 30° C. Koji preparation is then performed byfermenting the soybean meal with a low moisture content of 40 wt % bymeans of koji mold until the enzymes required in order to convert theproteins, isoflavones, saponins and phytic acid, etc., contained in thesoybean meal into respective health-promoting components such aspeptide, isoflavone aglycones, saponin aglycones, myo-inositol andMaillard reaction products, etc., are produced.

In concrete terms, in the case of isoflavones and saponins, therespective glycosides are decomposed so that aglycones are produced.Furthermore, in the case of phytic acid, which is a compound in whichphosphate groups are bonded to all of the hydroxy groups ofmyo-inositol, the above-described phosphate groups are dissociated bymeans of an enzyme that breaks down phytic acid, so that one or more ofthe compounds inositol pentaphosphate, inositol tetraphosphate, inositoltriphosphate, inositol diphosphate, inositol monophosphate and inositolare produced.

With respect to such cases in which a health-promoting component isproduced by this koji preparation step, a case will be described inwhich the glycosides of isoflavones and saponins are decomposed so thatrespective isoflavone aglycones and saponin aglycones are produced.

In this case, an enzyme known as β-glucosidase, which decomposes theglycosides of isoflavone compounds and is manufactured by koji mold as aresult of the propagation of koji mold in the soybean meal, breaks downthe glycosides of the isoflavone compounds in the soybean meal and thusproduces isoflavone aglycones. Furthermore, an enzyme known asβ-glucuronidase, which decomposes the glycosides of saponins and ismanufactured by the koji mold, breaks down the glycosides of thesaponins contained in the soybean meal and thus produces saponinaglycones.

The koji mold used in this koji preparation is koji mold that have longbeen used in Japanese fermented food products and tempeh; for example,koji mold from the genus Aspergillus and genus Rhizopus which are stablein food products, such as Aspergillus usami, Aspergillus kawachii,Aspergillus awamori, Aspergillus saitoi, Aspergillus oryzae andAspergillus niger, etc. are desirable for use.

In regard to the fermentation time, the fermentation time is set atleast 24 hours or longer in accordance with the type of koji mold used.It is desirable that the fermentation time be sufficient to causethorough decomposition of the glycosides of the isoflavones and saponinsin the soybean meal.

The temperature of the mixture in this koji preparation apparatus variesover time as shown for example in FIG. 2 as the manufacture of kojiprogresses. In particular, from the initiation of koji preparation untila state of “mori” (the first agitation) is reached after 22 hours, thetemperature gradually rises. Then, when “mori” (the first agitation) ispassed, the temperature drops slightly, and then again gradually risesuntil a state of “naka” (the second agitation) is reached after 27hours. When the mixture in a state of “naka” (the second agitation) isagitated, the temperature drops slightly, and then again rises until astate of “shimai” (the final agitation) is reached after 32 hours. Whenthe mixture in a state of “shimai” (the final agitation) is agitated,the temperature drops slightly, and then again rises until a time of 40hours has elapsed. Afterward, the temperature gradually drops, and kojipreparation is completed after 48 hours.

Under the above-described koji preparation conditions, the lactic acidbacteria (with an intestine-regulating effect in single-stomachedanimals) that are added simultaneously with the inoculation of the kojimold in the above-described koji preparation step propagate veryfavorably; and the multiplication of other undesirable organisms can bereliably suppressed. Lactobacillus bulgaricus and Streptococcusthermophilus, etc., which are commonly used in yogurt in Europe, may beused as lactic acid bacterial agents.

Using microbiological nomenclature, Bifidobacterium bifidum,Bifidobacterium infantis, Bifidobacterium longum, Enterococcus faecalis,Lactobacillus acidophilus, Lactobacillus casei, Lactococcus lactis andPediococcus sp., etc., may be cited as examples of such lactic acidbacteria that have an intestine-regulating function in single-stomachedanimals.

The lactic acid bacteria used as such bacteria with anintestine-regulating function have good compatibility with the koji moldused in koji preparation, and show extremely good propagation themselveswithout hindering the propagation of the koji mold, so that theproduction efficiency is also superior.

Next, water is added to the product following the completion of kojipreparation, and the product is maintained for a prescribed time in astate in which the temperature is elevated to a temperature of 30 to 65°C.; accordingly, the glycosides of the isoflavones and saponinscontained in the soybean meal are sufficiently diminished by thedecomposing actions of the β-glucosidase and β-glucuronidase containedin the product. As a result, hydrolysis is performed while aglycones ofthe isoflavones and saponins are produced.

In regard to the hydrolysis of the above-described proteins, it isdesirable to use a hydrolysis time and hydrolysis temperature that aresufficient to cause a thorough diminishment of the glycosides of theisoflavones and saponins contained in the soybean meal in accordancewith the type of koji mold used.

If this is done, organic acids will be produced in the initial stage offermentation so that the propagation of undesirable organisms in thesoybean meal is suppressed. Accordingly, there is no danger of secondarycontamination, and a product using soybean meal as a raw material can beproduced in large quantities.

Table 4 shows the contents of isoflavone compounds in 100 g of soybeanmeal in a case where untreated soybean meal were subjected to kojipreparation for 48 hours at an initial temperature of 30° C., afterwhich an amount of water equal to the weight of the product was added,and protein hydrolysis was performed for an additional 24 hours at 30°C.

TABLE 4 Daidzin Daidzein Genistin Genistein 25 74 531 59 (Units: mg/100g)

According to this Table 4, the amounts of daidzein and genistein, whichare aglycones of isoflavone compounds, were 74 mg and 59 mg, values thatwere respectively approximately 23 times and 14 times the values seen inthe conventional example shown in FIG. 1; thus, the amounts of thesecompounds were greatly increased. It is seen from this that the amountsof daidzein and genistein produced can be further increased byperforming hydrolysis for 24 hours or longer following the completion ofkoji preparation.

With the same object as Table 4, Table 5 shows the values beforetreatment and after treatment for other embodiments in which thetreatment of the process of the present invention was performed onuntreated soybean meal and separated soybean protein.

Describing the soybean meal first, the ratio of koji mold to rawmaterial was selected so that 0.1 g of crudely refined white ricecontaining koji mold spores at the rate of 8×10⁷ spores/g was added to100 g of soybean meal constituting the raw material. Using this ratio ofkoji mold to raw material, untreated soybean meal was subjected to kojipreparation for 48 hours at an initial temperature of 30° C.; then, anamount of water equal to the weight of the product was added, andprotein hydrolysis was performed for an additional 48 hours at 50° C.The results obtained were as shown in Table 5.

Meanwhile, the above-described commercially marketed soybean proteinused was FUJINIC 200 (commercial name) manufactured by Fuji PurinaProtein Kabushiki Kaisha. In this case, the ratio of koji mold to rawmaterial was selected so that 0.1 g of crudely refined white ricecontaining koji mold spores at the rate of 8×10⁷ spores/g was added to100 g of commercially marketed soybean protein constituting the rawmaterial. Using this ratio of koji mold to raw material, untreatedcommercially marketed soybean protein was subjected to koji preparationfor 48 hours at an initial temperature of 30° C.; then, an amount ofwater equal to the weight of the product was added, and proteinhydrolysis was performed for an additional 48 hours at 50° C. Theresults obtained were as shown in Table 5.

TABLE 5 Commercially marketed soybean Soybean meal protein Before AfterBefore After Daidzin 100 Not detected 90 1.0 Daidzein 3.2 70 5.3 100Genistin 120 1.3 120 3.3 Genistein 4.2 64 4.4 94 (Units: mg/100 g)

According to this Table 5, the amounts of daidzein and genistein, whichare aglycones of isoflavone compounds, in the case of soybean meal, were70 mg and 64 mg, which were respectively approximately 22 times and 15times the values measured prior to treatment; thus, the amounts of thesecompounds were greatly increased. Furthermore, daidzin, which is anisoflavone compound with glycosides, was decomposed until it could nolonger be detected, and genistin was also greatly diminished to a valueof 1.3 mg.

Furthermore, in the case of commercially marketed soybean protein aswell, the amounts of daidzein and genistein, which are aglycones ofisoflavone compounds, were 100 mg and 94 mg, which were respectivelyapproximately 19 times and 21 times the values measured prior totreatment; thus, the amounts of these compounds were greatly increased.Furthermore, daidzin and genistin, which are isoflavone compounds withglycosides, were greatly diminished to 1.0 mg and 1.3 mg, respectively.

Thus, according to the present invention, aglycones which have a highpharmacological effect among isoflavones and saponins, etc. contained insoybeans can be manufactured with an extremely high productionefficiency.

Next, the manufacture of a product, in which free myo-inositol isproduced as a health-promoting component and phytic acid contained insoybean meal is removed, will be described.

In this case, enzymes known as phytase and phosphatadase which decomposephytic acid and are manufactured by koji mold as a result of thepropagation of such koji mold in soybean meal decompose and remove thephytic acid contained in soybean meal.

More specifically, from phytic acid, which is a compound in whichphosphate groups are bonded to all of the hydroxy groups ofmyo-inositol, the above-described phosphate groups are dissociated bymeans of an enzyme that breaks down phytic acid, so that one or more ofthe compounds inositol pentaphosphate, inositol tetraphosphate, inositoltriphosphate, inositol diphosphate, inositol monophosphate and inositolare produced, thus removing the phytic acid.

The koji mold used in this koji preparation is koji mold that have longbeen used in Japanese fermented food products and tempeh; for example,koji mold from the genus Aspergillus and genus Rhizopus which are safeas food products and which have a high phytase potency and phosphatadasepotency, such as Aspergillus usamii, Aspergillus kawachi, Aspergillusawamori, Aspergillus saitoi, Aspergillus oryzae and Aspergillus niger,etc., are desirable for use.

In regard to the fermentation time, the fermentation time is set atleast 24 hours or longer in accordance with the type of koji mold used;and it is desirable that the fermentation time be sufficient to causethorough removal of the phytic acid contained in the soybean meal.

In the subsequent hydrolysis process and decomposition process, water isadded to the product following the completion of koji preparation, andthe product is then maintained for a prescribed period of time in astate in which the product is heated to a temperature of 30 to 55 ° C.As a result, the phytic acid contained in the soybean meal issufficiently diminished by the decomposing actions of the phytase andphosphatadase contained in the product, and hydrolysis is performed.

In regard to this hydrolysis, it is desirable that the hydrolysis timeand hydrolysis temperature used be sufficient to cause thorough removalof the phytic acid contained in the soybean meal in accordance with thetype of koji mold used.

Furthermore, the removal of phytic acid is accomplished by liberating atleast one phosphate group from the phytic acid, which consists ofinositol hexaphosphate. The number of phosphate groups liberated may bedetermined in accordance with the desired health-promoting action. Inthis case, the number of phosphate groups liberated from the phytic acidmay be controlled by adjusting the fermentation time, hydrolysis timeand hydrolysis temperature in accordance with the type, state,characteristics and quantity of the beans, the type, state,characteristics and quantity of the koji mold, and the type andcharacteristics, etc. of the product.

Table 6 shows the phytic acid content in 100 g of soybean meal foruntreated soybean meal, soybean meal A and B which were subjected tokoji preparation for 48 hours at an initial temperature of 30° C. usingtwo types of shochu malt (Aspergillus niger and Aspergillus awamori),and which were then subjected to hydrolysis for an additional 24 hoursat 30° C. following the addition of an equal weight of water to therespective products, and soybean meal which was subjected to aconventional alcohol cleaning treatment.

TABLE 6 Object soybean meal Phytic acid content (mg/100 g) Untreatedsoybean meal 999 (mg/100 g) Shochu malt treatment A Not detected Shochumalt treatment B Not detected Alcohol-cleaned soybean meal 1150 (mg/100g) (Detection limit: 5 mg/100 g)

According to this Table 6, the phytic acid content in untreated soybeanmeal was 999 mg, or approximately 1%, while the phytic acid contents ofsoybean meal A and B, which were subjected to a shochu malt treatmentaccording to the process of the present invention, and which were thensubjected to protein hydrolysis for an additional 24 hours at 30° C.following the addition of an equal weight of water to the respectiveproducts, were diminished to a point where said phytic acid contentscould not be detected, i. e., to a point where all of the phytic acidwas decomposed.

On the other hand, the phytic acid content in the soybean meal subjectedto a conventional alcohol cleaning treatment was 1150 mg, and thusshowed no decrease at all.

Thus, according to the present invention, it is possible to achieve agreat reduction or almost complete elimination of phytic acid insoybeans.

Next, the propagation of lactic acid bacteria (which are a type ofintestine-regulating bacteria) in the present invention will bedescribed.

In the present embodiment, lactic acid bacteria are added simultaneouslywith the inoculation of the koji mold; accordingly, these lactic acidbacteria propagate in large quantities in the process leading up to thecompletion of hydrolysis. Furthermore, since lactic acid bacteria areacid-producing bacteria, the propagation of other undesirable organismscan be reliably suppressed by the acid produced; and the productobtained has a good fragrance and taste. Lactic acid bacteria which havean intestine-regulating action in single-stomached animals are containedin large quantities in the product thus produced, and the product alsocontains bacteriocin produced by the lactic acid bacteria; accordingly,an intestine-regulating action can be reliably manifested insingle-stomached animals when such animals ingest this product.

To describe this further, using defatted soybeans as a raw material,miso koji mold (Aspergillus oryzae) and lactic acid bacteria(Lactococcus lactis subsp. lactis) were mixed with the soybeans in suchproportions that 0.1 g of crudely refined white rice containing kojimold spores of the miso koji mold at the rate of 8×10⁷ spores/g wasadded to 100 g of the raw-material defatted soybeans, and the number oflactic acid bacteria added was on the order of 10³ colony forming unit(cfu)/g. Then, koji preparation was performed for 48 hours at an initialtemperature of 30° C., and protein hydrolysis was performed for anadditional 41 hours at 50° C. following the addition of an equal weightof water to the product. In this case, the number of lactic acidbacteria in respective products dried by a hot air draft immediatelyafter the completion of koji preparation (koji product), and following13 hours, 24 hours, 30 hours and 41 hours of hydrolysis, wasinvestigated. Furthermore, 0.1 g of each of the products was added to100 ml of milk and separately to 100 ml of a suspension prepared bydissolving 16 g of commercially marketed skim milk in 140 ml of water,and the pH was investigated immediately after addition and after 18hours at 40° C. The results obtained are shown in Table 7.

TABLE 7 Number of Milk Defatted powdered milk lactic acid After 0 After18 After 0 After 18 bacteria hours hours hours hours Koji product (1.2 ×6.77 4.73 6.65 4.80 10⁷) After 13 hours 3.4 × 10⁹ 6.78 4.54 6.63 4.54 ofhydrolysis After 24 hours 2.9 × 10⁹ 6.77 4.54 6.64 4.57 of hydrolysisAfter 30 hours 3.1 × 10⁹ 6.76 4.55 6.63 4.54 of hydrolysis After 41hours 2.2 × 10⁹ 6.76 4.52 6.63 4.55 of hydrolysis

According to this Table 7, the lactic acid bacteria were greatlyincreased from the order of 10³ colony forming unit (cfu)/g to the orderof 10⁷ colony forming unit (cfu)/g in the koji preparation step and werefurther greatly increased to the order of 10⁹ colony forming unit(cfu)/g in the subsequent hydrolysis process. Furthermore, it is alsoseen that the compatibility of the miso koji mold (Aspergillus oryzae)and the lactic acid bacteria (Lactococcus lactis subsp. lactis) is good.Moreover, since no ventilation was performed in the hydrolysis process,even the anaerobic lactic acid bacteria showed considerable growth. Whenthis concentration on the order of 10⁹ colony forming unit (cfu)/g wasreached, no undesirable organisms recognized at the time of completionof the koji preparation step could be confirmed. The amount of lacticacid following hydrolysis was increased to a sufficient number;furthermore, it may be said that the propagation of undesirable aerobicorganisms was reliably suppressed in the environment that was presentduring hydrolysis, and that the propagation of undesirable organisms wasalso reliably suppressed as a result of the action shown by Lactococcuslactis subsp. lactis in producing bacteriocin (see Gross, E. and Morell,J. L.: J. Am. Chem. Soc., 93, 4634-4635 (1971)). Furthermore, in themilk and skim milk suspensions containing 0.1 g of the respectivesamples, the pH was caused to drop from approximately 6.5 toapproximately 4.5 after 18 hours had passed (at 40° C.) from the time ofaddition; thus, it is seen that the lactic acid bacteria showed furtheractive propagation.

In regard to the timing of the addition of the lactic acid bacteria,these bacteria may be added at any time from the initiation of the kojipreparation treatment to a point just prior to the completion ofhydrolysis; however, it is desirable to insure a time that allows theprescribed propagation of the lactic acid bacteria.

In cases where the product containing a health-promoting componentmanufactured by completing a solid culture treatment in the mannerdescribed above is used in, for instance, food products, etc., themanufactured product can be used “as is” or can be formed as a finalproduct by being dried and pulverized as shown in FIG. 1. In the presentembodiment, lactic acid bacteria are used; accordingly, if these lacticacid bacteria are obligate anaerobes, it is desirable that the bacteriabe dried by a freeze-drying method so that the lactic acid bacteria arenot killed. On the other hand, in cases where the lactic acid bacteriaare facultative anaerobes, drying by means of hot air draft drying at alow drying temperature may be used besides the above-describedfreeze-drying method.

Next, the product of the present invention which contains ahealth-promoting component will be described.

The product of the present invention includes a direct product that usesbeans as raw materials and an applied product which uses such a directproduct as a raw material (inclusive of cases where such a directproduct is used as a portion of the raw material). For example suchproducts include food products, livestock feeds, pet foods and drug rawmaterials, etc.

The product according to the process of the present invention usingbeans as a raw material can be manufactured without adding salt;accordingly, a food product with an extremely low salt content isobtained, so that large quantities of this product can be eaten when theproduct is used as a foodstuff.

Furthermore, a health-promoting component such as peptides, isoflavoneaglycones, saponin aglycones, myo-inositol and Maillard reactionproducts, etc., as well as a health-promoting component consisting oflactic acid bacteria, which are one type of intestine-regulatingbacteria, and bacteriocin, which is a product of such bacteria, arecontained in such food products.

Among the above-described health-promoting component, the componentsother than lactic acid bacteria can be very easily absorbed by thedigestive tracts of single-stomached animals, etc. Furthermore, asdescribed above, these health-promoting components comprise at least onemember of the group consisting of liver function improving constituents,cardiac function improving constituents, anti-inflammatory constituents,antifat constituents, antioxidation consititents and antimutagenconstituents; accordingly, a diet exhibiting superior effects in termsof health maintenance in humans can be enjoyed.

Furthermore, the lactic acid bacteria included in the health-promotingcomponent have an intestine-regulating action in the intestines ofsingle-stomached animals; accordingly, in cases where theintestine-regulating function is already good, this intestine-regulatingfunction can be improved even further; and in cases where theintestine-regulating function is poor, this intestine-regulatingfunction can be recovered, thus contributing to the maintenance andpromotion of health in single-stomached animals.

For example, in regard to food products according to the product of thepresent invention, if this product is formed into configurations such asbiscuits or cakes, etc., which can easily be eaten by humans, or if theproduct is formed “as is” (without being heated) int( a drink such as ayogurt flavored drink or cocoa flavored drink, etc., which can easily beimbibed by humans, then, when the food product is eaten or imbibed, ahealth-promoting component which is superior in terms of a hepaticfunction strengthening action, a cardiac function strengthening action,an anti-inflammatory action, a slenderizing action, an antioxidantaction, an anti-mutagenic action and an intestine-regulating action,etc., can be ingested at the same time. In particular, if biscuits areeaten so that the daily ingestion requirement considered necessary inorder to obtain the above-described health-promoting actions issatisfied, such ingestion alone is useful in preventing illness.

According to the preparation process of the present invention, live kojimold are grown, and aglycones which have a high pharmacological effectamong isoflavones and saponins contained in beans are manufactured at anextremely high production rate; furthermore, the phytic acid containedin the beans is removed, and proteins are hydrolyzed. Therefore,aglycones can easily be produced and phytic acid can easily be removedin the case of solid beans; moreover, the manufacturing process issimple, and manufacturing costs are low.

Furthermore, in the preparation process of the present invention, aconventional koji preparation apparatus can be utilized “as is.” Thereis no particular need to manufacture production base equipment. Thus,the present invention has high all-purpose utility.

EXAMPLES

The health-promoting action of the product of the present invention willbe described below in terms of concrete practical examples ofapplication.

Example 1

In the present practical example, a comparison of intestine-regulatingaction was made between an ordinary feed for suckling piglets, and feedsprepared by adding fermented defatted soybeans manufactured by theprocess of the present invention or plasma protein to the same ordinaryfeed.

Experimental Methods

(1) Test Feeds

The respective feeds used were the base feed (control group) shown inTable 8 below, which was used as a feed for suckling piglets, a feed(test group) prepared by adding fermented defatted soybeans prepared bythe process of the present invention to the base feed at a rate of 4%,and a feed (comparative group) prepared by adding plasma protein to thesame base feed at a rate of 1.8%, with the crude protein contents beingadjusted so that said contents were equal.

TABLE 8 Indicated components of base feed and names of raw materials,etc. Feed for suckling piglets (used during artificial suckling of Typeof feed piglets) Indicated components Crude protein 22.0% or greaterCrude fat 3.0% or greater Crude fiber 2.0% or less Crude ash content6.5% or less Calcium 0.65% or greater Phosphorus 0.50% or greater TDN84.0% or greater DCP 20.0% or greater Names of raw materials, etc.Classification of raw materials Amount added Name of raw material¹⁾Grains 46% Powdered wheat, bread crumbs, dextrin, (soy- bean flour)²⁾Feeds consisting of animal 32% Skim milk, fish meal, substances (driedwhey) Vegetable oils and lees 3% Concentrated soybean protein, potatoprotein Other 19% Sugar, glucose, animal oils, plasma protein, breadyeast, calcium phosphate, bacillus toyoi bacteria, fructooligo-saccharides, silicic acid, citric acid, tartaric acid, lactic acid,malic acid, (calcium carbonate), (betaine), (salt) Feed additivesAvilamycin (good's name) 40 g potency/ton Morantel citrate 30 g/tonColistin sulfate 40 g potency/ton Notes) ¹⁾Raw material names are listedsubstantially in the order of amount added ²⁾Raw materials shown inparentheses may be omitted in some cases depending on the raw materialconditions, etc.

(2) Test Animals

26 WL×D hybrids (13 males and 13 females) were used as suckling piglets.The average body weight for all of the animals at the initiation oftesting was 8.0 kg, and the average age was approximately 26 days.

(3) Test Methods

Inside a concrete pig shed with a natural ventilation system, theanimals of the control group, test group and comparative group wereplaced in 190 cm×125 cm×75 cm all-drain board cages, with 6 animals toeach cage so that the body weights were equal. Each group was raised fortwo weeks with free feeding and watering allowed. Each week, the bodyweights of the animals and the amounts of feed ingested were measured,and the body weight increase and feed requirements were determined fromthese values. The results are shown in Table 9. The health conditions ofthe piglets were also observed daily.

TABLE 9 Body weight, weight increase, amount of feed ingested and feedrequirements. Comparative Numbers of Control group group Test groupanimals 6 6 4 Body weight (kg) After 0 days  8.3 ± 0.8 (100)  8.3 ± 0.9(100)  8.2 ± 0.8 (98) After 7 days 10.2 ± 1.3 (100) 10.4 ± 1.2 (103)10.7 ± 0.9 (105) After 14 days 14.3 ± 1.7 (100) 14.5 ± 1.5 (102) 15.5 ±1.0 (109) Increase in body weight (g/day/animal) After 0 to  267 ± 102(100)  300 ± 131 (112) 365 ± 88 (137) 7 days After 7 to 587 ± 73 (100)586 ± 68 (100) 688 ± 21 (117) 14 days After 0 to 427 ± 86 (100) 443 ± 91(104) 526 ± 42 (125) 14 days Amount of feed ingested (g/day/animal)After 0 to 366 (100) 376 (103) 398 (109) 7 days After 7 to 742 (100) 851(115) 991 (134) 14 days After 0 to 554 (100) 613 (111) 679 (123) 14 daysFeed requirement After 0 to 1.37 (100) 1.25 (92) 1.09 (80) 7 days After7 to 1.26 (100) 1.45 (115) 1.44 (114) 14 days After 0 to 1.30 (100) 1.39(107) 1.29 (99) 14 days Notes) -Mean value ± standard deviation (n − 1)-Numerals in parentheses are indices with the value for the controlgroup taken as 100.

There was no diarrhea in any of the groups, and no deficiencies in bodytone appeared. As shown in Table 9, the weight increase in the periodfrom 0 to 14 days, during which the health conditions of the pigletsappeared normal, was 25% higher in the test group than in the controlgroup. This tendency appeared in the first week following the initiationof testing, with the increase in body weight during the period of 0 to 7days being 37% higher in the test group than in the control group.However, this difference was not significant. The weight increase duringthe period of 7 to 14 days was 17% higher in the test group than in thecontrol group, and this difference was significant (p<0.05). In thecomparative group in which plasma protein was added, the weight increaseduring the period of 0 to 14 days was only 4% higher than in the controlgroup, so that no great effect was observed. Since no great increase inbody weight was observed even in the case of a feed prepared by addingordinary untreated defatted soybeans to the base feed, the resultssuggested that fermented defatted soybeans manufactured by the processof the present invention did not constitute a minus factor in terms ofdigestive absorption in young single-stomached animals such as piglets,etc., but rather showed a large plus factor, i. e., the presence of anintestine-regulating action.

Example 2

In this practical example, an experiment was conducted concerning theamount of a hepatic drug metabolizing enzyme (amount of cytochromeP-450) in respective feeds using fermented defatted soybeansmanufactured according to the process of the present invention,untreated defatted soybeans and casein as protein sources.

Experimental Methods

Male mice of std:ddy at an age of 4 weeks and with a body weight ofapproximately 20 g (according to Nippon SLC) were raised for 7 days on acommercially marketed solid feed (MF manufactured by Oriental Kobo(yeast) Kabushiki Kaisha) in order to acclimatize the animals to theraising environment. Afterward, the animals were divided into threegroups as shown in Table 10 below, i. e., a casein group (8 animals)using casein as a feed, a fermented defatted soybean group (8 animals)using fermented defatted soybeans manufactured by the process of thepresent invention as a feed, and an untreated defatted soybean group (8animals) using unfermented defatted soybeans as a feed. These animalswere raised for 4 weeks; on the final day of raising, the animals werekilled by decapitation, and the livers were excised.

TABLE 10 Compositions of test feeds. Fermented Untreated Casein defatteddefatted Component (g/kg) soybeans (g/kg) soybeans (g/kg) Casein 200 — —Fermented defatted — 325 — soybeans Untreated defatted soybeans — — 365Mineral mixture 35 35 35 Vitamin mixture 10 10 10 Corn oil 50 50 50Soybean fiber 16 — 8 Sucrose 200 200 200 α-cone starch 489 380 332

(1) Preparation of Hepatic Microsomes

The livers excised as described above were immersed in an ice-cooled0.15 M KCl solution, and excess blood was washed away. Afterward, a 0.1M tris-hydrochloric acid buffer A (pH 7.4) was added, and a homogenatewas prepared using a homogenizer. This homogenate was centrifugallyseparated (9000*G, 15 minutes, 4° C.), and the supernatant wascollected. This supernatant was again centriftigally separated(105,000*G, 90 minutes, 4° C.), and the pellets thus obtained weresuspended in a 0.1 M tris-hydrochloric acid buffer B (pH 7.4), thusproducing hepatic microsome samples.

(2) Measurement of Protein Concentration

Measurements were performed using bovine serum albumin as a standardaccording to the method of Lowry (O. H. Lowry et al.: J. Biol. Chem.,193, 265, 1951).

(3) Measurement of Amount of Cytochrome P-450

Measurements were performed using the Method of Omura and Sato (T.Omura, R. Sato, J. Biol. Chem., 239, 2370, 1964).

As shown in Table 11 below, the measurement results were as follows: i.e., the amount of cytochrome P-450 was significantly lowest in thecasein feed group. When the fermented defatted soybean group and theuntreated defatted soybean group were compared, an increase in theamount of cytochrome P-450 was observed in the fermented defattedsoybean group. It can be seen that this probably suggests that theamount of cytochrome P-450 in a dose-dependent manner with isoflavoneaglycones. In other words, if the three test feeds are compared, it isseen that the fermented defatted soybeans manufactured by the process ofthe present invention cause the greatest increase (with a significantdifference) in the amount of cytochrome P-450 (which is a hepatic drugmetabolizing enzyme). Furthermore, cytochrome P-450 II also contributesto the promotion of metabolism in Practical Examples 3 and 4 describedbelow; thus, it can be seen that fermented defatted soybean manufacturedby the process of the present invention cause a significant increase inthe promotion of metabolism.

TABLE 11 Effects of fermented defatted soybeans on microsome proteinlevels and cytochrome P-450 level in hepatic microsomes. Microsomeprotein Feed Liver weight (g) (mg/g liver) Casein 2.07 ± 0.08^(b) 23.0 ±1.0^(a) Fermented defatted soybeans 1.99 ± 0.11^(ab) 25.2 ± 1.1^(a)Untreated defatted soybeans 1.78 ± 0.08^(a) 26.1 ± 2.1^(a) CytochromeP-450 Cytochrome P-450 Feed (nmol/mg protein) (nmol/g liver) Casein 0.42± 0.04^(a)  9.7 ± 1.2^(a) Fermented defatted soybeans 0.92 ± 0.05^(c)23.4 ± 2.1^(c) Untreated defatted soybeans 0.65 ± 0.03^(b) 17.1 ±1.8^(b) Notes) -Mean value ± standard deviation (n = 8) -Mean values invertical rows which do not have the same characters show significantdifferences (P < 0.05) -Testing for significant differences wasaccomplished by Duncan's multiple range test.

Example 3

In the present practical example, a feed using unfermented defattedsoybeans as a raw material and a feed using fermented defatted soybeansmanufactured by the process of the present invention as a raw materialwere compared with respect to the rate of acetaldehyde metabolism (inconnection with the strengthening of hepatic function).

Experimental Methods

Using 15 ddy male mice with a body weight of approximately 20 g in eachtest group, the unfermented defatted soybean feed and the fermenteddefatted soybean feed which was manufactured by the process of thepresent invention, both shown in Table 12 below, were given to the micefor four weeks. Afterward, a 5% acetaldehyde suspension (suspended indistilled water) was injected into the abdominal cavity of each animalat the rate of 10 ml per kg of body weight. The time in a coma (from theonset of coma to re-awakening) was then measured.

TABLE 12 Experimental feeds Unfermented defatted Fermented defattedsoybean soybean feed (g/kg) feed (g/kg) Unfermented 402 — defattedsoybean feed Fermented defatted — 359 soybean feed Corn oil 50 50Vitamin mixture 10 10 Mineral mixture 35 35 Cellulose 12 — αcorn starch291 346 Sucrose 200 200

As shown in Table 13 below, the measurement results indicate that thecoma time was long, i. e., approximately 96 minutes, in the case of thegroup receiving the unfermented defatted soybean feed; on the otherhand, the coma time was approximately 71 minutes, shortened byapproximately 25 minutes, in the case of the group receiving thefermented defatted soybean feed manufactured by the process of thepresent invention. Thus, the product of the present invention canaccelerate the rate of acetaldehyde metabolism and is thus able toprevent hangovers and deleterious after-effects of alcohol, and tostrengthen hepatic function.

TABLE 13 Effect on coma time caused by acetaldehyde administration Comatime (minutes) Unfermented defatted soybean feed (n = 15) 96 ± 9^(a)Fermented defatted soybean feed (n = 15) 71 ± 7^(b)

Example 4

In the present practical example, on the basis of the effect ofshortening acetaldehyde coma time observed in the fermented defattedsoybeans in the above-described Practical Example 3, the effects ofvarious types of protein sources (including fermented defatted soybeansmanufactured by the process of the present invention) on acuteacetaldehyde toxicity were compared.

Experimental Methods

Using 14 ddy male mice with a body weight of approximately 20 g in eachgroup, the feeds shown in Table 14 below, i. e., an unfermented defattedsoybean feed, a fermented defatted soybean feed manufactured by theprocess of the present invention, casein, egg white and gluten, weregiven to the mice for four weeks. Afterward, a 10% acetaldehydesuspension (suspended in distilled water) was injected into theabdominal cavity of each animal at the rate of 10 ml per kg of bodyweight, and the survival rate was measured.

TABLE 14 Protein source in feed (survival rate) Fermented Untreateddefatted defatted soybeans soybeans Casein Egg white Gluten Same day11/14 10/14 8/14 5/14 2/14 (79%) (71%) (57%) (36%) (14%) First day 11/149/14 8/14 5/14 0/14 (79%) (64%) (57%) (36%) (0%) Second day 8/14 6/147/14 4/14 0/14 (57%) (43%) (50%) (29%) (0%) Third day 8/14 4/14 7/143/14 0/14 (57%) (29%) (50%) (21%) (0%) Fourth day 6/14 3/14 5/14 3/140/14 (43%) (21%) (36%) (21%) (0%) Fifth day 6/14 3/14 4/14 3/14 0/14(43%) (21%) (29%) (21%) (0%)

It was confirmed from these measurement results that mice fed with afermented defatted soybean feed manufactured by the process of thepresent invention as a protein source had a higher survival rate thanmice fed with an unfermented defatted soybean feed, casein, egg white orgluten as a protein source. Thus, it was confirmed that a capacity tobreak down acetaldehyde can be obtained by administering the fermenteddefatted soybean feed manufactured by the process of the presentinvention. In other words, it appears that there was an effect thatincreased the activity of aldehyde dehydrogenase, which is asecond-phase detoxifying enzyme. It appears that this function is anaction obtained by means of a product in which defatted soybeans aresubjected to koji preparation using koji mold and are then hydrolyzed,so that components in the soybeans are changed by some type of enzyme.

Example 5

In this practical example, an evaluation of antioxidant action wasperformed by means of an AOM test, using products manufactured by theprocess of the present invention along with other substances asspecimens. The results obtained are shown in FIG. 3.

Here, the products of the present invention that were used as specimenswere a 0.1% ethanol extract of fermented and extracted soybean protein,and a 0.5% ethanol extract of fermented and extracted soybean protein.Furthermore, the other specimens used were an untreated extractedsoybean protein (no additives), a 0.1% ethanol extract of untreatedextracted soybean protein, a 0.1% ethanol extract of koji mold, and a0.1% extract of 98% α-tocopherol.

The AOM test performed here is a test in which the respective specimensare added to lard, after which the resulting mixtures are heated to 90°C., and the peroxide value (POV) is measured over time.

It is seen from the results in FIG. 3 that the 0.1% ethanol extract offermented and extracted soybean protein and 0.5% ethanol extract offermented and extracted soybean protein constituting products of thepresent invention required a long time for the peroxide value (POV) tobecome elevated compared to untreated extracted soybean protein and werethus superior in terms of antioxidant action.

Thus, the product of the present invention has an antioxidant actionsimilar to that of α-tocopherol and can suppress the oxidation of oil bybeing added in small amounts to dishes using large amounts of oil, etc.Accordingly, the product of the present invention can prevent liverdamage caused by peroxidized lipids.

Example 6

The present practical example concerns anti-mutagenic action. Theprocess of koji preparation according to the present invention wasperformed on two types of raw materials, i. e., whole soybeans anddefatted soybeans; and respective products prepared by hot air draftdrying following koji preparation and hot air draft drying followinghydrolysis were subjected to a test in which the suppression rate of theSOS induction reaction of Salmonella typhimurium was measured by MNNG inorder to ascertain the respective anti-mutagenic properties.

TABLE 15 Suppression rate (%) Whole soybeans Untreated 22.5 Followingkoji preparation 15.2 (product dried by hot air draft) Followinghydrolysis 60.2 (product dried by hot air draft) Defatted Untreated 0.8soybeans Following koji preparation 24.5 (product dried by hot airdraft) Following hydrolysis 34.7 (product dried by hot air draft)

The measurement results were as shown in Table 15 above. The suppressionrate following hydrolysis tended to be highest; and it appears from thisthat Maillard reaction products were produced by hydrolysis. Thus, inthe present invention, a Maillard reaction occurs in the same manner asin miso; and there is no addition of salt as there is in miso or soysauce. Accordingly, the hydrolysis time following koji preparation canbe greatly shortened.

The present invention is by no means restricted to the above-describedembodiments and Examples, and various modifications may be made asnecessary.

What is claimed is:
 1. A product containing a health-promotingcomponent, said product consists of: a health-promoting componentproduced by inoculating koji mold on a pulse crop to effect kojipreparation, and then hydrolyzing products produced by said kojipreparation treatment; and a health-promoting component consisting ofbacteria which have an intestine-regulating effect in single-stomachedanimals and are added to said pulse crop and grown during a periodextending from an inoculation of said koji mold to a completion of saidhydrolysis.
 2. A product containing a health-promoting componentaccording to claim 1, wherein said bacteria which have anintestine-regulating effect in single-stomached animals are addedsimultaneously with said inoculation of said koji mold.
 3. A productcontaining a health-promoting component according to claim 1 or 2,wherein said bacteria which have an intestine-regulating effect insingle-stomached animals consist of lactic acid bacteria which have anintestine-regulating effect in single-stomached animals.
 4. A productcontaining a health-promoting component according to claim 1 or 2,wherein said health-promoting component produced by said kojipreparation treatment and hydrolysis treatment comprises at least onemember of the group consisting of liver function improving constituents,cardiac function improving constituents, anti-inflammatory constituents,antifat constituents, antioxidation consititents and antimutagenconstituents.
 5. A product containing a health-promoting componentaccording to claim 1 or 2, wherein said health-promoting componentproduced by said koji preparation treatment and hydrolysis treatment areformed so as to be absorbed by digestive tract.
 6. A product containinga health-promoting component according to claim 3, wherein saidhealth-promoting component produced by said koji preparation treatmentand hydrolysis treatment comprises at least one member of the groupconsisting of liver function improving constituents, cardiac functionimproving constituents, anti-inflammatory constituents, antifatconstituents, antioxidation consititents and antimutagen constituents.7. A product containing a health-promoting component according to claim3, wherein said health-promoting component produced by said kojipreparation treatment and hydrolysis treatment are formed so as to beabsorbed by digestive tract.
 8. A product containing a health-promotingcomponent according to claim 4, wherein said health-promoting componentproduced by said koji preparation treatment and hydrolysis treatment areformed so as to be absorbed by digestive tract.
 9. A product containinga health-promoting component according to claim 6, wherein saidhealth-promoting component produced by said koji preparation treatmentand hydrolysis treatment are formed so as to be absorbed by digestivetract.